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(Actinopterygii) in the State of Rio De Janeiro, Brazil

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(Actinopterygii) in the State of Rio De Janeiro, Brazil NEMATODE AND CESTODE LARVAE OF HYGIENIC-SANITARY IMPORTANCE IN Lopholatilus villarii (ACTINOPTERYGII) IN THE STATE OF RIO DE JANEIRO, BRAZIL Aline Monteiro da SILVA1,2, Marcelo KNOFF2*, Nilza Nunes FELIZARDO1, Delir Corrêa GOMES2, Sérgio Carmona de SÃO CLEMENTE1 ABSTRACT The tile fish, Lopholatilus villarii, occurs from Brazil to Argentina where it is commercially important it because possesses highly appreciated meat. The aim of this study was to identify the larvae of anisakid and raphidascaridid nematodes and trypanorhynch cestodes in L. villarii purchased in fish markets of the municipality of Niterói, state of Rio de Janeiro, Brazil, and calculate their parasitological indices and present their sites of infection. From August 2015 to September 2016, 31 specimens of L. villarii were investigated. The fish were necropsied and their viscera and musculature analyzed. The helminths found were processed according to standard helminthological techniques for taxonomic identification. Of 31 fish analyzed, 28 (90.3%) were parasitized with larvae of anisakid and raphidascaridid nematodes, Anisakis sp., Terranova sp., Pseudoterranova sp. and Hysterothylacium deardorffoverstreetorum (L3 and L4) and plerocercus of the trypanorhynch cestode Otobothrium sp. Their parasitological indices were calculated. The helminths parasitized mainly the serosas of intestine, stomach and liver, and H. deardorffoverstreetorum was also parasitizing the abdominal musculature of one fish. This is the first report of anisakid and raphidascaridid nematodes and trypanorhynch cestodes in L. villarii. Hygienic-sanitary aspects were discussed. Key words: Malacanthidae; Anisakidae; Raphidascarididae; Trypanorhyncha. LARVAS DE NEMATOIDES E CESTOIDES DE IMPORTÂNCIA HIGIÊNICO-SANITÁRIA EM Lopholatilus villarii (ACTINOPTERYGII) NO ESTADO DO RIO DE JANEIRO, BRASIL RESUMO O peixe batata, Lopholatilus villarii, ocorre do Brasil a Argentina e possui uma carne apreciada e comercialmente importante. O objetivo deste estudo foi identificar as larvas de nematoides Anisakidae e Raphidascarididae e cestoides Trypanorhyncha de L. villarii obtidos em mercados de pescado do município de Niterói, Estado do Rio de Janeiro, Brasil, calcular seus índices parasitários e apresentar os sítios de infecção. De agosto de 2015 a setembro de 2016, foram investigados 31 espécimes de L. villarii. Os peixes foram necropsiados e suas vísceras e musculatura e analisados. Os helmintos encontrados foram processados de acordo com as técnicas em helmintologia para permitir a identificação taxonômica. Dos 31 peixes analisados, 28 (90,3%) estavam parasitados com larvas de nematoides anisaquídeos e rafidascaridídeos: Anisakis sp., Terranova sp., Pseudoterranova sp. e Hysterothylacium deardorffoverstreetorum (L3 e L4) e plerocercos de cestoides Trypanorhyncha: Otobothrium sp. Seus índices parasitários foram calculados. Os helmintos estavam parasitando, principalmente, as serosas do intestino, estômago e fígado e H. deardorffoverstreetorum estava também na musculatura abdominal de um peixe. Este é o primeiro registro de nematoides anisaquídeos e rafidascaridídeos e cestoides Trypanorhyncha em L. villarii. Aspectos higiênico- sanitários foram discutidos. Palavras-chave: Malacanthidae; Anisakidae; Raphidascarididae; Trypanorhyncha. Artigo Científico: Recebido em 14/03/2017; Aprovado em 30/06/2017 1Laboratório de Inspeção e Tecnologia de Pescado, Faculdade de Medicina Veterinária, Universidade Federal Fluminense, Rua Vital Brazil Filho, 64, Vital Brazil, 24320-340 Niterói, RJ, Brasil. 2Laboratório de Helmintos Parasitos de Vertebrados, Instituto Oswaldo Cruz, FIOCRUZ, Av. Brasil, 4365, Manguinhos, 21040-900 Rio de Janeiro, RJ, Brasil. E-mail: [email protected] (corresponding author) B. Inst. Pesca, São Paulo, 43(3): 385 - 398, 2017 Doi: 10.20950/1678-2305.2017v43n3p385 386 SILVA et al. INTRODUCTION hypersensitivity has been demonstrated for some species in studies using a murine model (RODERO The tile fish, Lopholatilus villarii Miranda-Ribeiro, and CUÉLLAR, 1999; VÁZQUEZ-LÓPEZ et al., 2001, 2002; 1915 (Malacanthidae) occurs in the Southwest GÒMEZ-MORALES et al., 2008; MATTOS et al., 2013). Atlantic from Brazil to Argentina, and is known The aim of this study was to report, for the for its good tasting meat, which has commonly first time, larvae of anisakid and raphidascaridid been marketed fraudulently as mottled grouper, nematodes and trypanorhynch cestodes as parasites Mycteroperca rubra Bioch, 1793, or Argentinian of L. villarii marketed in the municipality of Niterói, sandperch, Pseudopercis semifasciata Cuvier, 1829 RJ, Brazil. Parasitological indices were calculated (FIGUEIREDO and MENEZES 1980; LIMA and and infection sites presented, with comments on MESQUITA, 1996; PAIVA and ANDRADE-TUBINO, the hygienic-sanitary significance of these parasites. 1998; FROESE and PAULY, 2016). The nematodes of the families Anisakidae and Raphidascarididae have been reported parasitizing METHODS several Brazilian marine fish in studies on classical and molecular taxonomy, ecology and hygiene and From August 2015 to September 2016, 31 health, since the larvae migrate after the death of the specimens [total length 43-71 (52.7) cm; weight hosts (FELIZARDO et al., 2009; DIAS et al., 2010,2011; 0.970 – 4.810 (1.925) kg] were acquired from fish KNOFF et al., 2012,2013; FONTENELLE et al., markets in the municipality of Niterói, state of Rio 2013,2015; RIBEIRO et al., 2014; PANTOJA et al., 2015; de Janeiro, Brazil. The fish were transported in an KURAIEM et al., 2016; FONSECA et al., 2016). Human isothermal box to the laboratory for necropsy, where infections with these nematodes have been reported they were identified according to FIGUEIREDO and in Europe (Netherlands, Spain, France, Italy, Belgium, MENEZES (1980). Denmark, England and Germany), in the Americas After necropsy, the internal organs were (United States of America, Chile, Peru and Brazil), in transferred to Petri dishes containing physiological Korea and, especially, in Japan, where more than 2,000 solution with 0.65% NaCl. The nematode larvae cases occur annually (ADAMS et al., 1997; AUDICANA found dead were fixed in AFA (70% alcohol - et al., 2002; ACHA and SZYFRES, 2003; CHAI et al., formaldehyde - acetic acid), while live larvae 2005; CRUZ et al., 2010; D’AMICO et al., 2014). were fixed in hot AFA at 60 ºC, so they would die Cestodes of the order Trypanorhyncha are of distended, preserved in a solution of 70 °GL ethanol hygienic importance because of their repugnant plus 5% glycerin and clarified with Amman’s aspect, particularly when teleostean fish present lactophenol (KNOFF and GOMES, 2012). Taxonomic massive infections in their musculature and organs classification of nematodes was in accordance with that may make commercialization infeasible PETTER and MAILLARD (1988); TIMI et al. (2001); due to sanitary inspection and/or rejected by the FELIZARDO et al. (2009); KNOFF et al. (2012) and consumer (AMATO et al., 1990; SÃO CLEMENTE FONSECA et al. (2016). et al., 2004,2007; FELIZARDO et al., 2010). This Plerocerci of trypanorhynchs were transferred order possesses a great diversity of species with to distilled water and the cysts opened under a global distributions. As adults, these worms inhabit stereomicroscope with the aid of sharp needles to the stomach and intestine of elasmobranch fish, release the larvae, which were then refrigerated for while the larval forms can infect a large number of at least 24h to permit the relaxation of scolices and teleostean fish and marine invertebrates in tropical tentacular extroversion. The larvae were then fixed in and subtropical regions, but rarely freshwater fish cold AFA, stained with Langeron´s carmine, clarified and other vertebrates (CAMPBELL and BEVERIDGE, in beechwood creosote, and preserved as whole 1994; PALM, 2004). These cestodes cause significant mounts on Canada balsam according to KNOFF and economic losses. Furthermore, accidental infections GOMES (2012). The classification of Trypanorhyncha of humans by larval trypanorhynchs have been followed PALM (2004), BEVERIDGE and JUSTINE reported due to the ingestion of raw fish meat, and (2007) and SCHAEFFNER and BEVERIDGE (2013). although they do not present zoonotic potential, Measurements were obtained by bright field recent research reports that these cestodes can cause microscopy using an Olympus BX 41 microscope. allergic disorders in humans because immunological The samples were then analyzed by bright-field B. Inst. Pesca, São Paulo, 43(3): 385 - 398, 2017 Nematode and cestode larvae of hygienic-sanitary importance... 387 microscopy with a Zeiss Axiophot microscope Maldonado Jr, Torres, Pinto and Gomes, 2012 using Nomarski’s differential interference contrast (Figures 6a-c; 7a-c)], and one fourth instar larvae (DIC) apparatus, and images obtained with a Canon (L4) [H. deardorffoverstreetorum (Figures 8a-c; 9a-c)]. digital camera (Power Shot A640). Some specimens Additionally, 26 fish (83.9%) were parasitized with a were prepared for scanning electron microscopy total of 414 trypanorhynch cestode plerocerci of the (SEM) as described by LOPES TORRES et al. (2013). family Otobothriidae, identified as Otobothrium sp. The samples fixed in 70% ethanol were dehydrated (Figures 10a-b; 11). in an ethanol series (70º to 100º GL), CO2 critical The morphological and morphometric data of point dried, coated in gold, and then examined and the helminths found are shown in Tables 1, 2 and 3. photographed using a SEM (Jeol JSM-6390LV), under The parasitological
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